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Method for preparing hydroxyapatite/silk fibroin compound porous stand material

A technology of hydroxyapatite and silk fibroin, applied in medical science, prosthesis, etc., can solve the problems of high brittleness, insufficient mechanical properties, and limited use of hydroxyapatite, and achieve less reagents, cheap reagents, and high performance. excellent effect

Inactive Publication Date: 2012-08-08
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, single-component hydroxyapatite has high brittleness, poor toughness, insufficient mechanical properties, slow degradation and easy migration from the implantation site, which limits its use as a human implant.

Method used

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  • Method for preparing hydroxyapatite/silk fibroin compound porous stand material
  • Method for preparing hydroxyapatite/silk fibroin compound porous stand material
  • Method for preparing hydroxyapatite/silk fibroin compound porous stand material

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preparation example Construction

[0032] 2. Preparation of 5% silk fibroin solution by weight concentration:

[0033] (1) the fibrous silk fibroin after degumming is 50% CaCl in weight percent concentration 2 In the solution, the weight ratio is 1:25, the temperature is 98° C., and the time is 5 minutes. After dissolving, filter and dialyze to obtain a silk fibroin solution with a weight concentration of 5%.

[0034] 3. Preparation of hydroxyapatite homogenate:

[0035] (1) 700ml of 60°C saturated Ca(OH) 2 The solution was placed in a water bath at 60°C, and 10% wt of H was slowly added dropwise 3 PO 4 Solution 11ml, during the dropping process, use a magnetic stirrer to continuously and rapidly stir and maintain the water temperature. The digital display pH meter monitors the pH of the solution. When the pH of the solution is lower than 7.0, add ammonia water dropwise to it, and adjust the pH of the solution to maintain at 8.7;

[0036] (2) After stirring for 3 hours, place it at 25°C for 48 hours, pour o...

Embodiment 1

[0038] (1) Accurately weigh a certain mass of hydroxyapatite homogenate, stir continuously with a magnetic stirrer and keep the water temperature at 25°C, and slowly add silk fibroin solution dropwise therein. The ratio of HA to SF was 1:9.

[0039] (2) Put the mixed solution of hydroxyapatite and silk fibroin into an ultrasonic cleaner, and vibrate for 3-5 minutes.

[0040] (3) Take out the mixed solution, slowly dropwise add glutaraldehyde crosslinking agent (commercially available) with a final concentration of 0.03% therein, and stir slowly with a magnetic stirrer evenly.

[0041] (4) Quickly transfer the mixed solution into a 24-well cell culture plate, and immediately put it into a -20°C refrigerator for 12 hours to pre-freeze.

[0042] (5) Take out the sample-loaded cell culture plate after freezing for 12 hours, return to temperature for 10 minutes at 25°C, and then put it back into the -20°C refrigerator for another 24 hours.

[0043] (6) Take out the sample-loaded ...

Embodiment 2

[0045] (1) Accurately weigh a certain mass of hydroxyapatite homogenate, stir continuously with a magnetic stirrer and keep the water temperature at 25°C, and slowly add silk fibroin solution dropwise therein. Make the ratio of HA to SF 3:7.

[0046] (2) Put the mixed solution of hydroxyapatite and silk fibroin into an ultrasonic cleaner, and vibrate for 3-5 minutes.

[0047] (3) Take out the mixed solution, slowly add a glutaraldehyde crosslinking agent with a final concentration of 0.03% dropwise therein, and stir slowly with a magnetic stirrer evenly.

[0048] (4) Quickly transfer the mixed solution into a 24-well cell culture plate, and immediately put it into a -20°C refrigerator for 12 hours to pre-freeze.

[0049] (5) Take out the sample-loaded cell culture plate after freezing for 12 hours, return to temperature for 20 minutes at 25°C, and then put it back into the -20°C refrigerator for another 24 hours.

[0050] (6) Take out the sample-loaded cell culture plate aft...

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Abstract

The invention discloses a method for preparing hydroxylapatite / fibroin protein composite porous support material, comprising the following steps: dropping fibroin protein solution in hydroxylapatite homogenate; putting the mixed solution in a supersonic cleaner for oscillation treatment; dropping glutaric dialdehyde cross linking agent to the mixed solution and stirring the mixture; and then transferring the solution to a cell culture plate and putting the solution into a refrigerator for precooling at minus 20 degrees; cooling the solution for 12h, refreezing the solution for 24h after temperature returning treatment at the temperature of 25 DEG C; then drying the solution again to prepare the hydroxylapatite / fibroin protein composite porous support material of different proportions. Thehydroxylapatite / fibroin protein composite multiaperture support material of the invention features simple preparation technique, short time, easily satisfied preparation conditions, no addition of pore-foaming agent, small use amount of reagent, low price and low cost thereof; in addition, the support material prepared in the invention features relatively high porosity factor, uniform dispersion of pores and mutual perforation of pores, thus meeting demands of bone tissue engineering support material on pore space and pore diameter structure.

Description

technical field [0001] The invention relates to the fields of biomaterial preparation and silk protein processing and utilization, in particular to a preparation method of a hydroxyapatite / silk fibroin composite porous scaffold material for bone tissue engineering. Background technique [0002] Bone injury and bone defect are common diseases in clinical practice, often require bone grafting treatment, the traditional treatment method is autologous or allogeneic bone grafting for bone repair. Autologous bone grafts have a good repair effect, but they have the disadvantages of limited supply and the need for a second operation. Allogeneic bone grafts have the disadvantages of immunogenicity and the possibility of spreading infectious diseases, so they are not ideal bone repair materials. The basic idea of ​​bone tissue engineering is to use a small amount of tissue cells to repair bone defects after in vitro culture and expansion. At the same time, it can be arbitrarily shaped...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/46A61L27/56A61L27/22A61L27/12
Inventor 朱良均丰强闵思佳张海萍
Owner ZHEJIANG UNIV
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